Efficient-secretion canine parvovirus resistant monoclonal antibody hybridoma cell A135 strain

A canine parvovirus, monoclonal antibody technology, applied in the direction of antiviral agents, antiviral immunoglobulins, antibodies, etc., can solve the problems of infecting sick dogs, neutralizing titers (low antiviral activity, unstable quality, etc. , to achieve a wide range of effects with excellent biological activity

Inactive Publication Date: 2015-09-02
JIANGSU ACAD OF AGRI SCI
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Problems solved by technology

At present, the anti-CPV immune serum prepared from native animals or heterologous animals in the market has unstable quality, difficult quality control, high production costs, and there is a huge risk of exogenous virus introduction and infection of sick dogs, resulting in serious Biosafety issues; the application of monoclonal antibody therapy can reduce the above-mentioned risks and improve the curative effect, but the existing anti-CPV monoclonal antibodies not only have low neutralization titer (antiviral activity), but also have low neutralization potency to different CPV subtypes. Potency varies widely, no neutralizing activity against some CPV subtype strains

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0018] (1) Establishment of monoclonal antibody hybridoma cell lines

[0019] 1. Preparation of CPV Antigen The virulent canine parvovirus CPV-JS12 strain isolated from CPVD immunized dogs (Li Yuehua, et al. Molecular characteristics of VP2 gene of canine parvovirus JS12 strain and its expression in Escherichia coli. Chinese Veterinary Science, 2013, 43(10): 991-998) were inoculated in FK81 cells (purchased from Nanjing Tianbang Biotechnology Co., Ltd.), the diseased cell culture was frozen and thawed three times, and the cell debris was removed by centrifugation. The discontinuous sucrose density gradient method was used for purification, the virus concentration was measured by an ultraviolet spectrophotometer, and the CPV immune antigen was prepared and stored at -20°C for future use.

[0020] animal immunity The purified CPV immune antigen was used to immunize female BALB / c mice aged 6-8 weeks (purchased from the Experimental Center of Comparative Medicine of Yangzhou...

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Abstract

The invention relates to an efficient-secretion canine parvovirus (CPV) resistant monoclonal antibody hybridoma cell A135 strain and belongs to the technical field of biology. The hybridoma cell A135 strain selected from the secretion CPV resistant monoclonal antibody hybridoma cell bank is excellent in biological performance and is injected with ascetic fluid generated in a BALB / C mouse peritoneal, so that neutralizing titer is up to 1010. In addition, the hybridoma cell A135 strain has moderate neutralizing capacity for various CPV subtype virus strain such as CPV-2a, CPV-2b, CPV-2c (a) and CPV-2c (b) and CPV strains from foxes and raccoon dogs, is wide in anti-CPV strain range and is used for clinical treatment of attacked dogs with CPV diseases, and effective rate is up to 100%.

Description

technical field [0001] The invention relates to a hybridoma cell highly secreting anti-canine parvovirus (CPV) monoclonal antibody, belonging to the field of biotechnology. Background technique [0002] Canine parvovirus disease (CPVD) is a canine viral infectious disease caused by canine parvovirus (CPV), characterized by hemorrhagic enteritis or non-suppurative myocarditis. The disease is highly contagious , The mortality rate is high, dogs of all ages can be infected, the mortality rate of puppies is as high as 70%, and the economic loss caused by the disease is huge. Although CPVD vaccination has been widely adopted for susceptible dogs, CPV is still one of the most serious infectious diseases that endanger the dog industry in my country because of its strong resistance to the outside world and easy mutation. [0003] CPV (also known as CPV-2) is a new type of parvovirus isolated from the stool of dogs with hemorrhagic enteritis by American scholars in 1977. Virus (min...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/08A61K39/395A61P31/20
Inventor 王永山夏兴霞毕振威李月华诸玉梅欧阳伟潘群兴王晓丽董晨红梅永杰王晶宇吴红玲
Owner JIANGSU ACAD OF AGRI SCI
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